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The Impact of Intermittent Hypoxic Training on Rat Glioma Experimental Model

Author: LiAiSong
Tutor: ZhuYuHong
School:
Course: Neurology
Keywords: glioma intermittent hypoxia Ki67 p53 vWF
CLC: R739.4
Type: Master's thesis
Year: 2012
Downloads: 13
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Abstract


Objective To investigate the impact of intermittent hypoxic stimulation on rat glioma experimental model.Methods Methods:Surgical microscope and stereotaxic apparatus were applied to produce rat C6glioma model.64male Wistar rats were randomly divided into5groups:group A:the control group rat right caudate nuclear injection of lOμl of DMEM (Dulbecc’s modified eagle medium, in DMEM) medium, were not hypoxic stimulation (n=8). Group B:hypoxic control group for the rat right caudate nucleus injection lOμlDMEM medium to hypoxic stimulation (n=8). Group C:glioma hypoxia group for the rat right caudate nuclear injection lOμl suspension containing1x106C6cells to hypoxic stimulation (n=20). Group D:glioma group for the right caudate nucleus of rats injected lOμl suspension containing1x106C6cells were not hypoxic stimulation (n=18). Group E:Hypoxic preconditioning of the glioma group, advance the hypoxic preconditioning in three weeks, and then injected lOμl suspension containing1x106C6cells in the rat right caudate nucleus (n=10). One week after modeling, MRI Magnetic resonance imaging (MRI) shows tumor in brains of rats in Group C, Group D and Group E. Group B and group C, were placed in the hypoxic chamber for concentration of8%O2inhalation1hours a day. Four groups of rats (ie, intermittent hypoxic stimulation two weeks after modeling to three weeks later) were randomly selected half of the MRI examination observation of tumor growth and tumor volume measurement, and immediately drawn. The remaining rats were six weeks after the model (ie, five weeks after the intermittent hypoxic stimulus) line MRI examination and drawn. Observed and recorded the survival status of the rats. Take different times in each group rat brain specimens of brain tissue HE staining, glial fibrillary acidic protein (of astrocytes of fibrillary Any irregularity in protein and GFAP), Ki67, of p53, von Willebrand factor (vWF) immunohistochemistry.Results l.Ki67expression in rat C6glioma model accepted intermittent hypoxic stimulation2week’s hypoxic stimulation later, Ki67expression were proved of statistically difference between group B and C (P<0.05). After5week’s hypoxic stimulation, difference of Ki67expression were found of having statistically significance between group B and C (P<0.05).Ki67expression in week2and5were found of having statistically difference in glioma groups (P<0.05). Since the transplant of tumor cells, the expression of Ki67in week3was higher than it in week6.2. p53expression in rat C6glioma model accepted intermittent hypoxic stimulation:After2weeks’hypoxic stimulation, p53expression were found of having statistically difference between group B and C (P<0.05), group C and E (P<0.05).After5weeks’hypoxic stimulation, p53expression were found of having statistically difference between group B and C (P<0.05).3.vWF expression in the rat C6glioma model accepted intermittent hypoxic stimulation:After2weeks’hypoxic stimulation, vWF expression were of found of having statistically difference between group B and C (P<0.05).After5weeks’hypoxic stimulation, vWF expression were found of having statistically difference between gourp B and C (P<0.05).vWF expression were found of statistically different after hypoxic stimulation for2weeks and5weeks in group C (P<0.05), while such difference of vWF expression (P<0.05) was also found in group D at same time sets. After the transplant of C6glioma cells, vWF expression in week3was higher than it in week6.Conclusion1. C6cells were growthgrowed steadily, and were able to meet the experimental requirements.2. All Wistar rats injected C6cell were suffered from glioma, and the model is stable.3. The hypoxia program using of hypoxic chambers was easy to control and repeat. 4. In this study, the expression of Ki67,p53, and vWF in a rat model were no significant statistical difference between group C and other control groups. Hypoxia time extended to five weeks, only vWF was found of having significant difference compared to two weeks of intermittent hypoxic stimulation.5. Intermittent hypoxic stimulation did notneither increase the tumor cell proliferation, did not nor increasethe accumulation of mutant p53protein. It have had no obvious protective effect for the rat glioma model, and no adverse effects.

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